A well-defined biological product prepared by the The ... · linked glycan due to a higher...

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The European Biosimilar

Experience

Professor Paul Declerck

Laboratory for Therapeutic and Diagnostic Antibodies

[email protected] 2016 11 28 [email protected]

Biological medicinal product

A well-defined biological product prepared by the use of living systems, such as organisms, tissue

cultures or cells.

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2016 11 28 [email protected]

• Glycosylation

• Phosphorylation

• Sulfation

• Methylation

• N-acylation

• S-Nitrosylation

• ….

• cell type and culture

conditions

Molecular basis of heterogeneity

• Deamidation (e.g. Asn to Asp)

• Racemization (L to D)

• Oxidation ( Met, Tyr, His, Trp)

• Disulfide exchange

• …..

• External conditions (pH,

additives, temperature....)

> 108 variants


Infliximab, InFlixImab

cDNA infliximab

cDNA infliximab

InFlIxImab, inflIXimaB

cDNA infliximab

inflIXiMaB, Infliximab

The process determines the product

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McCamish. MAbs. 2011;3(2):209-17

Develop a highly

similar product

Concept of biosimilar development


Investigate reference product quality attributes to understand target variability, create boundaries within which to be “similar”

(Different lots examined over time)

Identify genetic information that codes for desired primary amino-acid sequence and clone into host cell using vector

(Same primary amino acid sequence)(Different cell line may be used)

Iterative process used to identify clones, producing protein with target posttranslational modifications

Converge to target product profile by iterative optimization of process


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McCamish. MAbs. 2011;3(2):209-17

Develop a highly

similar product

Confirm

Biosimilarity



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• Drug substance

• Manufacture

• Characterisation

• Control

• Reference standard

• Container

• Stability

• Drug product

• Description

• Development

• Manufacture

• Control

• Reference standard

• Container

• Stability

• Pharmacology

• Primary pharm.

• Secondary pharm.

• Safety pharm.

• Interactions

• Pharmacokinetics

• ADME

• Interactions

• Toxicology

• Single dose

• Repeat dose

• Genotoxicity

• Carcinogenicity

• Reproduction

• Local tolerance

• Pharmacology


• Single dose

• Repeat dose

• Special populations

• Efficacy and safety

• Dose finding

• Schedule finding

• Pivotal

• Indication 1

• Indication 2

• Indication 3

• Indication 4

• Post-marketing studies

Quality Nonclinical Clinical

Registration requirements (Original)

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9

• Drug substance

• Manufacture

• Characterisation

• Control• Reference standard

• Container

• Stability

• Drug product• Description

• Development

• Manufacture

• Control• Reference standard

• Container

• Stability

• Comparability data• Analytical

comparison with reference product

• Pharmacology

• Primary pharm.

• Secondary pharm.

• Safety pharm.• Interactions


• ADME

• Interactions

• Toxicology• Single dose

• Repeat dose

• Genotoxicity

• Carcinogenicity• Reproduction

• Local tolerance

• Pharmacology


• Single dose

• Repeat dose

• Special populations

• Efficacy and safety

• Dose finding

• Schedule finding

• Pivotal• Indication 1

• Indication 2

• Indication 3

• Indication 4

• Post-marketing studies

• Safety in larger population

• Efficacy in other indications

• Immunogenicity

Quality Nonclinical Clinical

Stu

dy

#1

Stu

dy

#2

Registration requirements (Biosimilar)


Registration of biosimilars (Europe)

• 2 refused by the EU commission:

o Interferon alpha-2a (2006)

o Insulin human (2015)

• 6 withdrawn:

o Insulin (2008)

• Insulin Rapid

• Insulin Long

• Insulin 30/70 Mix

o Insulin (2012)• Solumarv

• Isomarv medium

• Combimarv

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Solumarv (human insulin)

From European Public Assesment Report Solumarv®

Why refused?

• Insufficient details on manufacturing process

• Insufficiently demonstrated whether clinical study

batches are representative for market batches

• Insufficiently shown that quality of proposedbiosimilar is comparable to the reference product


Alpheon (Interferon alfa-2a)

From European Public Assesment Report Alpheon®

Why refused?

• Differences with reference product (e.g. impurities)

• Not enough data on stability

• Inadequate validation of process to make the finished

drug product

• Lower efficacy

• More side effects• Inadequately validated test to evaluate the potential to

trigger an immunological response

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• 28 approved in Europa (11/2016)

o 2 Human growth hormone (2006)

o 3 Epoietin alfa (2007)

o 2 Epoietin zeta (2007)

o 9 Filgrastim (2008 (4), 2009 (2), 2010, 2013, 2014)

o 3 Infliximab (2013 (2), 2016)

o 2 Follitropin alfa (2013, 2014)

o 2 Insulin glargine (2014, 2016)

o 1 Etanercept (2016)

o 2 Enoxaparin (2016)

o 2 Teriparatide (2016)




• 16 under review (11/2016)

o 2 Etanercept

o 2 Rituximab

o 4 Pegfilgrastim

o 3 Adalimumab

o 1 Insulin glargine

o 3 Trastuzumab

o 1 Insulin Lispro

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Physicochemical Comparability Tests:

Analytics Set the Foundation

Test Method Compares...

Amino acid analysis Amino acid composition

Peptide mapping

(LC-MS) in

combination with

MS/MS

Peptide coverage and chemical

modifications

Peptide mapping

(HPLC)

Tryptic peptide map by visual

inspection

N-terminal

sequencing

N-terminal sequences

C-terminal

sequencing

C-terminal sequences

Reduced mass Molecular weights by mass

spectrometry

Disulfide bonds Disulfide bonds location

Free thiol analysis Amount of free sulfhydryl groups

FTIR Secondary structures

CD Secondary structure

DSC Thermal stability; also determines

thermal transition temperatures

Test Method Compares…

Purity/Impurity

SEC-HPLC Aggregate content and monomeric

purity

CE-SDS

(reduced/nonreduced)

Electrophoretic mobility and purity

under nonreducing and reducingconditions

Charged Isoforms

IEF Isoelectric point(s)

IEC-HPLC Charge variant distribution

Glycosylation

Sialic acid analysis Sialic acid content

Monosaccharide

analysis

Neutral and amino sugar

composition

Oligosaccharide

profiling

Glycosylation pattern (eg,G0F,

G1F, G2F)

N-linked glycan

analysis

Oligosaccharide structures,

attachment sites, and distribution

Content

UV280Protein concentration

ELISA API content


• Binding to target

• Binding to

o FcɣRI, FcɣRII, FcɣRIII

o FcRn

o C1q

• Fab-associated functions (neutralization, activation, ...)

• Fc-associated functions (ADCC, CDC, complement activation, ...)

Biochemical and functional Comparability

Tests

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Biosimilar ESA (*)

• “Differences were observed at the glycosylation level”

• “Phosphorylated high mannose type structures were detected at higher levels than in ReferenceESA”

• “Lower values on N-glycolyl-neuramic acid and diacetylatedneuramic acids as compared to Reference ESA”

• “Peptide map showed differences … in O-linked glycan due to a higher sialylation and lower content of the oxidized variant”

Biosimilar IFX (*)

• “….. all major physicochemical characteristics and biological activities of biosimilar IFX were comparable to those of the reference product”

• “….difference in the amount of afucosylated infliximab, translating into a lower binding affinity towards FcγRIIIareceptors and a lower ex vivo antibody-dependent cellular cytotoxicity (ADCC) activity….”

• “… less intact IgG …. , mainly due to a higher proportion of non-assembled form. …. unlikely to impact its biological activity”

• “a higher level of C-terminal lysine variability”

• “…slightly higher level of aggregates …”

Biosimilars are Similar, not identical

(*) Based upon European Public Assessment Report on respective biosimilars.

How similar are biosimilars ?Biosimilar hGH (*)

• “The results of this study … demonstrate that Biosimilar rhGH produced at full scale is comparable to Reference Product”

• “The impurity profile of Biosimilar hGH shares some similarity with ReferencehGH; however the profiles are not identical”

• “ … impurities, … , are present in the Biosimilar hGH batches and are not in any Reference hGHbatches”

• “Additionally, there appears to be a higher level of deamidated variants in the Biosimilar hGH samples”


CT-P13 versus Infliximab reference

From European Public Assesment Report Remsima®

How similar are biosimilars ?

• Lower degree of afucosylation

• Lower binding affinity to FcƔRIIIa

• Lower activity in the most sensitive ADCC assay

• “However, no difference could be detected in several

experiments that are more representative of

pathophysiological conditions, and therefore more relevant clinically.”

Physicochemical / pre-clinical

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SB2 versus Infliximab reference

From European Public Assesment Report Flixabi®


• Lower degree of C-Lys variants ( CHO versus SP2/0 host cell)

• Lower % of charged variants

• Higher % of High Molecular Weight variants

• Higher binding affinity to FcƔRIIIa (114-141% vs. 77-

108%) but without impact on ADCC assay



From European Public Assesment Report Flixabi®


• Higher incidence of ADA formation in patients (47 % vs.38 % at day 71)

• Impact of ADA on efficacy is not clear (CHMP: Divergent opinion 14/36 negative)

• Data from studies in presence of MTX extrapolation of

immunogenicity to other indications?

SB2 versus Infliximab reference

Immunological events

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2016 11 28 [email protected] 2016 11 28 [email protected]

From European Public Assesment Report Benepali®


• Lower degree of C-Lys variants ( CHO versus SP2/0 host cell ?)

• Differences in charged variants

• Higher degree of afucosylation (considered not clinicallyrelevant because not involved in Mode of Action)

• Slightly less effective in a mouse model of arthritis (not

confirmed in clinical studies)

SB4 versus Etanercept reference


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From European Public Assesment Report Benepali®


• Significantly lower incidence of ADA formation in patients (overall 1 % vs. 13 % at wk 52)

• Reanalysis excluding wk 4 and 8: 0.3 vs. 0.7%

• Impact of assay methodology –low drug tolerance: data

affected by trough levels, the latter were different at wk4/8, thus reanalysis after excluding ADA data at wk 4/8

• “…. it is premature to conclude that SB4 is less immunogenic than reference ….”

SB4 versus Etanercept reference

Immunological events


From European Public Assesment Report Ovaleap® and Bemfola®

Study XM17 Study Afolia

Reference

(n=145)

XM17

(n=152)

Reference

(n=123)

Afolia

(n=249)

Ovarian

Hyperstimulation

Syndrome (OHSS)

2.7% 4.6% 13 % 32 %


“….. The following parameters [dose, oestradiol,body weight, age] were

comparable….. and cannot explain the difference that is observed in OHSS. The

observed difference could therefore be a chance finding. “(European Public Assesment Report Ovaleap® )

XM17 and Afolia versus FSH reference

Adverse events

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= BA B

Ref

Chemical drugs Biological drugs

B? ?A B

Ref


Originator biologic

One biosimilar

Two biosimilars

Four biosimilars

Biosimilars of originator R

originator R

biosimilar B1

biosimilar B2

biosimilar B3

biosimilar B4

Patients Could Face Multiple Switches Between Biosimilars to the Same Originator

Adapted from NIBRT ©

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• Substitution/switching is contraindicated

• Physician control over prescribing

• Appropriate (brand)naming required

• International pharmacovigilance systems should be imposed that enable unambiguous identification of the product associated with an adverse event

Biological drugs


Conclusions

• The concept for biosimilar development is well-defined

• The process for approval is rigorous

• Pharmaceutical quality of approved biosimilar is

guaranteed

• Differences in quality attributes are always present

• Major challenges include the identification of the

potential clinical relevance of differences in quality

attributes and non-clinical properties

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Conclusions

• Residual uncertainties (scientifically or statistically) have

so far always been deemed to have no impact on safety

and efficacy

• EPARs contain heterogenous information not consistent

between different biosimilars for the same reference

product

• To date, (multiple) switching between a reference and its

respective biosimilars can not be recommended since no

solid scientific data are available

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